MutT homologue 1 (MTH1) removes N6-methyl-dATP from the dNTP pool

MutT homologue 1 (MTH1) removes oxidized nucleotides from the nucleotide pool and thereby prevents their incorporation into the genome and thereby reduces genotoxicity. We previously reported that MTH1 is an efficient catalyst of O6-methyl-dGTP hydrolysis suggesting that MTH1 may also sanitize the nucleotide pool from other methylated nucleotides. We here show that MTH1 efficiently catalyzes the hydrolysis of N6-methyl-dATP to N6-methyl-dAMP and further report that N6-methylation of dATP drastically increases the MTH1 activity. We also observed MTH1 activity with N6-methyl-ATP, albeit at a lower level. We show that N6-methyl-dATP is incorporated into DNA in vivo, as indicated by increased N6-methyl-dA DNA levels in embryos developed from MTH1 knock-out zebrafish eggs microinjected with N6-methyl-dATP compared with noninjected embryos. N6-methyl-dATP activity is present in MTH1 homologues from distantly related vertebrates, suggesting evolutionary conservation and indicating that this activity is important. Of note, N6-methyl-dATP activity is unique to MTH1 among related NUDIX hydrolases. Moreover, we present the structure of N6-methyl-dAMP–bound human MTH1, revealing that the N6-methyl group is accommodated within a hydrophobic active-site sub-pocket explaining why N6-methyl-dATP is a good MTH1 substrate. N6-methylation of DNA and RNA has been reported to have epigenetic roles and to affect mRNA metabolism. We propose that MTH1 acts in concert with adenosine deaminase–like protein isoform 1 (ADAL1) to prevent incorporation of N6-methyl-(d)ATP into DNA and RNA. This would hinder potential dysregulation of epigenetic control and RNA metabolism via conversion of N6-methyl-(d)ATP to N6-methyl-(d)AMP, followed by ADAL1 catalyzed deamination producing (d)IMP that can enter the nucleotide salvage pathway

Kinetic and structural characterization of NUDT15 and NUDT18 as catalysts of isoprene pyrophosphate hydrolysis

Isoprene pyrophosphates play a crucial role in the synthesis of a diverse array of essential nonsterol and sterol biomolecules and serve as substrates for posttranslational isoprenylation of proteins, enabling specific anchoring to cellular membranes. Hydrolysis of isoprene pyrophosphates would be a means to modulate their levels, downstream products, and protein isoprenylation. While NUDIX hydrolases from plants have been described to catalyze the hydrolysis of isoprene pyrophosphates, homologous enzymes with this function in animals have not yet been reported. In this study, we screened an extensive panel of human NUDIX hydrolases for activity in hydrolyzing isoprene pyrophosphates. We found that human nucleotide triphosphate diphosphatase NUDT15 and 8-oxo-dGDP phosphatase NUDT18 efficiently catalyze the hydrolysis of several physiologically relevant isoprene pyrophosphates. Notably, we demonstrate that geranyl pyrophosphate is an excellent substrate for NUDT18, with a catalytic efficiency of 2.1 × 105 m−1·s−1, thus making it the best substrate identified for NUDT18 to date. Similarly, geranyl pyrophosphate proved to be the best isoprene pyrophosphate substrate for NUDT15, with a catalytic efficiency of 4.0 × 104 M−1·s−1. LC–MS analysis of NUDT15 and NUDT18 catalyzed isoprene pyrophosphate hydrolysis revealed the generation of the corresponding monophosphates and inorganic phosphate. Furthermore, we solved the crystal structure of NUDT15 in complex with the hydrolysis product geranyl phosphate at a resolution of 1.70 Å. This structure revealed that the active site nicely accommodates the hydrophobic isoprenoid moiety and helped identify key binding residues. Our findings imply that isoprene pyrophosphates are endogenous substrates of NUDT15 and NUDT18, suggesting they are involved in animal isoprene pyrophosphate metabolism

Diferencial de mortalidade em homens e mulheres em localidade da região Sudeste, Brasil: 1960, 1970 e 1980 The sex differential in mortality a city of Southeastern Brazil: 1960, 1970, 1980

Foram comparadas as taxas de mortalidade masculinas e femininas no Município do Rio de Janeiro (Brasil), em 1960, 1970 e 1980, buscando analisar os diferentes riscos a que homens e mulheres são submetidos, em cada grupo etário. Os diferenciais de mortalidade por sexo e causa foram estudados através da Razão de Sobremortalidade Masculina, das diferenças relativas e absolutas entre taxas e de taxas padronizadas. Além disso, foi desenvolvida uma análise dos diferenciais por grupos selecionados de causas para o ano de 1980. As taxas de mortalidade masculinas foram maiores do que as femininas em todas as faixas etárias, nos três anos estudados, com aumento da Razão de Sobremortalidade Masculina entre 15 e 34 anos , no período considerado. O excesso de mortes masculinas foi causado sobretudo pela elevação dos óbitos por causas violentas em homens jovens, o que retrata uma realidade dramática, cuja possibilidade de transformação através de medidas "técnicas" é escassa. Em relação aos óbitos por outras causas, os determinantes biológicos e os diferentes riscos a que são submetidos homens e mulheres devem ser considerados, buscando-se melhor compreensão da realidade.<br>Male and female mortality rates in the city of Rio de Janeiro in 1960, 1970 and 1980 are studied with a view to analysing the different risks to which men and women are subject by age group. Mortality differentials by sex and cause were studied by means of male/female mortality ratios, relative and absolute differences among rates, and standardized rates. An analysis of differentials by selected groups of cause for 1980 was undertaken. Male mortality rates were higher than the female rates in all age groups in the three years studied, with an increase of the male/female mortality ratio for the 15-34 age group over this period. The excess of male death was mainly due to the increase of deaths from violent causes among young men, a dramatic circunstance that can hardly be changed by "technical" procedures. As regards deaths from other causes, biological determinants and the different risks for men and women should be considered in order better to understand this situation